环保透明脱蜡液Van-Clear与传统试剂在组织处理后HE染色及FISH法检测乳腺癌HER2基因中的比较

发布时间：2018-04-24 20:40

本文选题：环保透明脱蜡液Van-Clear + 二甲苯　；参考：《华中科技大学学报(医学版)》2017年04期

【摘要】：目的比较环保透明脱蜡液Van-Clear和二甲苯透明脱蜡制作的组织切片苏木精-伊红(HE)染色优良率,对比荧光原位杂交(FISH)法检测2种透明脱蜡液制作的乳腺癌组织切片的人类表皮生长因子受体2(HER2)基因扩增阳性率,探讨Van-Clear替代二甲苯的可行性。方法收集中山市博爱医院2013年2月~2015年12月门诊及住院部送检的乳腺浸润性导管癌标本89例、乳腺增生标本125例、子宫平滑肌瘤标本193例、子宫平滑肌肉瘤标本12例、肺癌穿刺标本16例、绒毛标本139例,同一病变部位切取2个样本,用抽签法随机分为2组,命名为A、B组。A组采用二甲苯透明脱蜡制作切片574张;B采用Van-Clear透明脱蜡制作切片574张。依据切片染色情况判定切片等级,采用SPSS20.0软件比较A、B组切片HE染色优良率;A、B组中乳腺浸润性导管癌标本再各制作切片89张,采用SPSS 20.0软件比较FISH法检测的HER2基因扩增的差异。结果 (1)生物显微镜下,A、B两组切片HE染色结果细胞轮廓清晰、透明度好,细胞核与细胞质蓝红相映、色彩鲜艳,核质对比明显,核膜及核染色质颗粒清晰可见、分辨良好,分裂象染色体呈现黑蓝色,各种成分层次分明。(2)A、B两组切片HE染色优良片分别为570张、572张,中等、差片4张、2张。染色优良率分别为99.30%、99.65%,两组间染色优良率差异无统计学意义(χ2=0.50,P0.05)。(3)荧光显微镜下,A、B两组切片乳腺浸润性导管癌HER2双探针FISH检测结果组织轮廓和背景均清晰,探针定位准确,可见耀眼的红/绿荧光信号。(4)A、B两组切片检测乳腺浸润性导管癌HER2基因,FISH阳性率分别为24.72%、26.97%,两组间阳性率差异无统计学意义(χ2=0.50,P0.05),且FISH结果符合率为97.75%。结论环保透明脱蜡液Van-Clear有替代传统试剂二甲苯应用于组织切片HE染色及FISH法检测乳腺癌HER2基因的潜在可能。
[Abstract]:Objective to compare the excellent and good rate of Van-Clear and xylene transparent dewaxing solution in the staining of hematoxylin and eosin. The positive rate of human epidermal growth factor receptor 2H ER2 gene amplification was detected by fluorescence in situ hybridization (fish) method in two kinds of transparent dewaxing solution of breast cancer, and the feasibility of replacing xylene with Van-Clear was discussed. Methods from February 2013 to December 2015, 89 breast invasive ductal carcinoma specimens, 125 breast hyperplasia specimens, 193 uterine leiomyoma specimens and 12 uterine leiomyosarcoma specimens were collected from Boai Hospital, Zhongshan City. 16 cases of lung cancer and 139 cases of chorionic villi were divided randomly into two groups by drawing lots. The group A was named as group A and group A was made of 574 pieces of sections by transparent dewaxing of xylene. Van-Clear transparent dewaxing was used to make 574 pieces of sections. According to the staining of sections, the grade of sections was determined, and the excellent and good rate of HE staining was compared by SPSS20.0 software. 89 samples of invasive ductal carcinoma of breast were made in group A and B, and the difference of HER2 gene amplification detected by FISH method was compared with SPSS 20.0 software. Results 1) the results of HE staining showed that the cells in the two groups were clear in outline, clear in transparency, bright in color, bright in color, obvious in contrast of nuclear and cytoplasm, clearly visible in nuclear membrane and chromatin particles, and well distinguishable. The chromosomes of the mitotic elephant were dark blue, with 572 pieces of fine sections stained by HE, 572 pieces of medium size and 2 pieces of bad slices in HE stained sections of the two groups. The excellent and good staining rates were 99.30% and 99.65%, respectively. There was no significant difference between the two groups (蠂 ~ (2 +) 0.50) (P 0.05). The results of HER2 double-probe FISH detection of breast infiltrating ductal carcinoma in two groups under fluorescence microscope were clear, the tissue profile and background were clear, and the probe location was accurate. It can be seen that the positive rates of HER2 gene fish in the two groups were 24.72and 26.97, respectively. There was no significant difference between the two groups (蠂 ~ 2 ~ (0.50) P _ (0.05), and the coincidence rate of FISH results was 97.75). Conclusion Environmental transparent dewaxing solution Van-Clear has the potential to replace the traditional reagent xylene in HE staining of tissue sections and detection of HER2 gene in breast cancer by FISH method.
【作者单位】：南方医科大学附属中山博爱医院病理科;广东医科大学广东省医学分子诊断重点实验室;
【基金】：国家自然科学基金资助项目(No.81572782) 广东省医学科研基金立项课题(No.A2017321) 广东省中山市卫生和计划生育局医学科研立项课题(No.2014J128)
【分类号】：R737.9

【参考文献】